全文获取类型
收费全文 | 49680篇 |
免费 | 2861篇 |
国内免费 | 1719篇 |
专业分类
54260篇 |
出版年
2024年 | 110篇 |
2023年 | 741篇 |
2022年 | 974篇 |
2021年 | 1259篇 |
2020年 | 1326篇 |
2019年 | 1770篇 |
2018年 | 1623篇 |
2017年 | 1195篇 |
2016年 | 1247篇 |
2015年 | 1331篇 |
2014年 | 2488篇 |
2013年 | 3157篇 |
2012年 | 1818篇 |
2011年 | 2612篇 |
2010年 | 2750篇 |
2009年 | 2427篇 |
2008年 | 2342篇 |
2007年 | 2559篇 |
2006年 | 2188篇 |
2005年 | 2349篇 |
2004年 | 2138篇 |
2003年 | 1702篇 |
2002年 | 1367篇 |
2001年 | 1014篇 |
2000年 | 846篇 |
1999年 | 912篇 |
1998年 | 819篇 |
1997年 | 740篇 |
1996年 | 726篇 |
1995年 | 757篇 |
1994年 | 702篇 |
1993年 | 677篇 |
1992年 | 621篇 |
1991年 | 523篇 |
1990年 | 452篇 |
1989年 | 405篇 |
1988年 | 386篇 |
1987年 | 308篇 |
1986年 | 298篇 |
1985年 | 288篇 |
1984年 | 457篇 |
1983年 | 288篇 |
1982年 | 327篇 |
1981年 | 286篇 |
1980年 | 263篇 |
1979年 | 216篇 |
1978年 | 115篇 |
1977年 | 110篇 |
1976年 | 92篇 |
1975年 | 43篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
81.
Biserka Mulac-Jericevic Taghi Manshouri Tsuyoshi Yokoi M. Zouhair Atassi 《Journal of Protein Chemistry》1988,7(2):173-177
A set of 18 synthetic uniform overlapping peptides spanning the entire extracellular part (residues 1–210) of the -subunit of human acetylcholine receptor were studied for their binding activity of125I-labeled -bungarotoxin and cobratoxin. A major toxin-binding region was found to reside within peptide 122–138. In addition, low-binding activities were obtained with peptides 34–49 and 194–210. It is concluded that the region within residues 122–138 constitutes a universal major toxin-binding region for acetylcholine receptor of various species. 相似文献
82.
The Na+-independent binding of [3H]-alanine to rat brain stem plus spinal cord was reinvestigated, in order to study in more detail the characteristics of previously described -alanine binding processes. Binding was absent when amino acid-free postnuclear supernatants or crude synaptic membranes were used. Experiments performed with several other Na+-free preparations showed a sole binding component, irrespective of the preparation used. Biochemical characterization of this Na+-independent binding, using frozen/thawed/washed synaptosomal-mitochodrial fractions, showed that binding reached a plateau between 7 min and 13 min, increasing thereafter. Binding was linear with fraction protein over a range of 200–415 g/ml incubation medium. Binding was completely inhibited by glycine, alanine, -aminobutyric acid, -aminoisobutyric acid, hypotaurine and strychnine, and to a lesser extent by 2,2-dimethyl--alanine, brucine and gelsemine. It was insensitive to taurine, -aminobutyric acid (GABA), 2-guanidinoethanesulfonic acid (GES), carnosine, and bicuculline methiodide. Binding was reversible, saturable (K
D 20 M), and heat sensitive. 相似文献
83.
The proteolytic specificity of chicken cathepsin L was studied using bovine -casein as substrate. The peptide mixtures obtained after various times of hydrolysis were separated by RP-HPLC and ten peptides were identified. Chicken cathepsin L accepts proline residues in all positions except P
1
. Looking at the amino acid residues on the amino side of the scissile bond we found three times the Tyr-Pro pair at P
1
–P
2
positions and that the S
1
subsite can interact with modified amino acids such as phosphoserine.Abbreviations RP-HPLC
reverse phase high performance liquid chromatography
- NMec
N-methyl coumarylamide
- TEA
triethylamine
- TFA
trifluoroacetic acid 相似文献
84.
A number of proteases have been immobilized on alumina in a two-step procedure: the first step converted them into semisynthetic phosphoproteins which, in the second step, spontaneously bonded to alumina through their phosphate function. The immobilized enzymes thus obtained showed the physical properties typical of the inorganic carrier and a high activity on low molecular weight substrates. 相似文献
85.
A model is proposed for the effect of gramicidin A on the order and structure of phospholipid dispersions. According to this model, the addition of gramicidin A influences the surrounding lipids via two independent mechanisms. The first arises from a drop in surface pressure for those lipids substantially bounded by gramicidin A. The second mechanism arises from the increase in the phospholipid headgroup spacing due to the small polar region of the polypeptide. The model provides an explanation for the currently available NMR, X-ray diffraction and Langmuir monolayer results. The model also suggests mechanisms for the ability of gramicidin A to trigger a transition of the lipid from the lamellar to hexagonal II phase, the dependence of this transition on the lipid chain length and the formation of a lamellar phase with lysophosphatidylcholine.Abbreviations NMR
nuclear magnetic resonance
- DMPC
dimyristoylphosphatidylcholine
- S
molecular order parameter
- CSA
chemical shift anisotropy
- DPPC
dipalmitoylphosphati-dylcholine
- LPC
lysophosphatidylcholine 相似文献
86.
A. M. Cashmore M. S. Albury C. Hadfield P. A. Meacock 《Molecular & general genetics : MGG》1988,212(3):426-431
Summary The yeast 2 m circle encodes four major transcribed open reading frames, A, B, C and D. Products of ORF's A, B and C, together with the inverted repeats and the other cis-acting loci ORI and STB, have been shown to be involved in plasmid maintenance. However, the function of ORF D has remained unclear. We have therefore carried out studies on 2 m derivatives with both insertional and frameshift mutations in D. Our results indicate that there is a protein product encoded by ORF D, which is involved in plasmid maintenance. When the copy number of the C gene was reduced to one, by chromosomal integration, we observed striking differences in the efficiency of partitioning of D
+ and D
– plasmid derivatives. Absence of D function could be compensated by an increase in dosage of the C gene, indicating that the D product may act to regulate C expression. Since the C product has been implicated in copy number control as well as partitioning, our data suggest that the D product may also be involved in both of these processes. 相似文献
87.
Summary Stage V and VI (Dumont, J.N., 1972.J. Morphol.
136:153–180) oocytes ofXenopus laevis were treated with collagenase to remove follicular cells and were placed in K-free solution for 2 to 4 days to elevate internal [Na]. Na/K pump activity was studied by restoring the eggs to normal 3mm K Barth's solution and measuring membrane current-voltage (I–V) relationships before and after the addition of 10 m dihydroouabain (DHO) using a two-microelectrode voltage clamp. Two pulse protocols were used to measure membraneI–V relationships, both allowing membrane currents to be determined twice at each of a series of membrane potentials: (i) a down-up-down sequence of 5 mV, 1-sec stair steps and (ii) a similar sequence of 1-sec voltage pulses but with consecutive pulses separated by 4-sec recovery periods at the holding potential (–40 mV). The resulting membraneI–V relationships determined both before and during exposure to DHO showed significant hysteresis between the first and second current measurements at each voltage. DHO difference curves also usually showed hysteresis indicating that DHO caused a change in a component of current that varied with time. Since, by definition, the steady-state Na/K pumpI–V relationship must be free of hysteresis, the presence of hysteresis in DHO differenceI–V curves can be used as a criterion for excluding such data from consideration as a valid measure of the Na/K pumpI–V relationship. DHO differenceI–V relationships that did not show hysteresis were sigmoid functions of membrane potential when measured in normal (90mm) external Na solution. The Na/K pump current magnitude saturated near 0 mV at a value of 1.0–1.5 A cm–2, without evidence of negative slope conductance for potentials up to +55 mV. The Na/K pump current magnitude in Na-free external solution was approximately voltage independent. Since these forward-going Na/K pumpI–V relationships do not show a region of negative slope over the voltage range –110 to +55 mV, it is not necessary to postulate the existence of more than one voltage-dependent step in the reaction cycle of the forward-going Na/K pump. 相似文献
88.
Expression sites of genes encoding (13,14)--glucan 4-glucanohydrolase (EC 3.2.1.73) have been mapped in germinated barley grains (Hordeum vulgare L.) by hybridization histochemistry. A32P-labelled cDNA (copy DNA) probe was hybridized to cryosections of intact barley grains to localize complementary mRNAs. No mRNA encoding (13,14)--glucanase is detected in ungerminated grain. Expression of (13,14)--glucanase genes is first detected in the scutellum after 1 d and is confined to the epithelial layer. At this stage, no expression is apparent in the aleurone. After 2 d, levels of (13,14)--glucanase mRNA decrease in the scutellar epithelium but increase in the aleurone. In the aleurone layer, induction of (13,14)--glucanase gene expression, as measured by mRNA accumulation, progresses from the proximal to distal end of the grain as a front moving away from, and parallel to, the face of the scutellum.Abbreviations cDNA
copy DNA
- RNase
ribonuclease 相似文献
89.
Terry L. Riss Kenneth P. Karey B. Daniel Burleigh David Farker David A. Sirbasku 《In vitro cellular & developmental biology. Plant》1988,24(11):1099-1106
Summary A serum-free clonal density growth assay was developed for the quantification of the biological activity of human recombinant
insulin-like growth factor I (IGF-I). The assay measures IGF-I stimulated growth of Balb/c 3T3 cells cultured over 4 d on
poly-d-lysine-coated plastic surfaces in a serum-free medium formulation composed of a 1∶1 (vol/vol) mixture of Ham's F12 and Dulbecco's
modified Eagle's media, supplemented with 3.0 ng/ml bovine basic fibroblast growth factor (bFGF), 10 μg/ml human transferrin,
100 μg/ml ovalbumin, and 1.0 μM dexamethanose. Low-temperature trypsinization of serum-supplemented stock cultures combined with the use of poly-d-lysine-coated plates made it unnecessary to use serum or fibronectin to promote cell attachment and survival. Serum-free
growth conditions were optimized with respect to the concentrations of the supplements. Addition of IGF-I resulted in 3.5-fold
more cells than control cultures without IGF-I after 4 d. Deletion of bFGF resulted in no IGF-I stimulation of growth. The
concentrations of various preparations of IGF-I required to achieve one-half maximal stimulation of cell number (ED50), ranged between 1.25 and 4.7 ng/ml. In parallel assays, IGF-I was 6.6 times more potent than human recombinant insulin-like
growth factor II and 32 times more potent than insulin. When cells were seeded into medium containing IGF-I, transferrin,
ovalbumin, and dexamethasone but no bFGF, growth was minimal. Dose-response addition of bFGF showed an ED50, of 0.9 ng/ml. The methods reported are useful to monitor the biological potency of recombinant and natural-source growth
factors as well as providing a new means of studying the multiple growth factor requirements of Balb/c 3T3 cells in cultures.
This work was supported by a contract from IMCERA Bioproducts, Inc. 相似文献
90.
A soluble Lewis blood-group gene associated -3/4-L-fucosyltransferase has been purified from human milk by a series of steps involving hydrophobic chromatography on Phenyl Sepharose 4B, ion exchange chromatography on CM-Sephadex C-50, affinity chromatography on GDP-hexanolamine Sepharose 4B and gel filtration on Sephacryl S-200. The first step separated -3-L-fucosyltransferase activity directed towardsN-acetylglucosamine in Type 2 (Gal1-4GlcNAc-R) acceptors from an -3/4-fucosyltransferase fraction acting on both Type 1 (Gal1-3GlcNAc-R) and Type 2 acceptors. Further purification of this latter fraction on CM-Sephadex and GDP-hexanolamine Sepharose gave a single peak of fucosyltransferase activity that catalysed the addition of fucose toN-acetylglucosamine in both Type 1 and Type 2 acceptors and to theO-3 position of glucose in lactose-based oligosaccharides. The enzyme preparation at this stage resembled previously described -3/4-fucosyltransferase preparations purified from human milk. However, gel filtration of this preparation on Sephacryl S-200 or Sephadex G-150 separated further amounts of -3-fucosyltransferase activity acting solely on Type 2 acceptors and left a residual -3/4-fucosyltransferase that retained strong -4 activity with the Type 1 acceptor, lacto-N-biose 1, and -3 activity with 2-fucosyllactose, but had relatively little -3 activity withN-acetyllactosamine and virtually no capacity to transfer fucose to glycoproteins withN-linked oligosaccharide chains having unsubstituted terminal Type 2 structures. 相似文献